目的探讨EB病毒潜伏性膜蛋白1(LMP1)羧基末端第三活性区在鼻咽上皮细胞生长中的作用。方法采用逆病毒感染的方法,建立稳定表达野生型LMP1(LMP1^WT)和突变型LMP1(LMP1^△232-351)的鼻咽上皮细胞(NP69)系,然后通过细胞生长曲线、平皿克隆形成与软琼脂集落实验、细胞周期与抗凋亡检测等方法,观察LMP1羧基末端第三活性区对鼻咽上皮细胞生物学特性的影响。结果LMP1^△232-351体外促转化细胞生长能力较LMP1^WT明显降低(P〈0.01);NP69-LMP1^△232-351细胞的凋亡与细胞G1期分布均较NP69-LMP1^WT细胞增加(P〈0.01)。结论LMP1羧基末端第三活性区可能通过调节细胞周期与凋亡而影响鼻咽上皮生物学特性。
Objective To investigate the biological character of nasopharyngeal epithelial cells affected by carboxy terminal activating region - 3 of the Epstein - Barr virus encoded latent membrane protein 1 ( LMP1 ). Methods We adopted the method of retroviruses infection and established the nasopharyngeal epithelial cell lines ( NP69 ) of stable expression wild type LMP1 ( LMP1^WT) and mutant LMP1 (LMP1^△232-351 ). Then, we made use of cell growth curve, plate clone formation, soft agar colony experiment, cell cycle and anti - apoptosis detection to observe biological character of nasopharyngeal epithelial cell affected by carboxy terminal activating region -3 of LMP1. Results Compared with LMP1^WT, the growth ability, in vitro, of LMP1^△232-351 transformation cells obviously decreased (P 〈0.01 ). The numbers of cell apoptosis and cell proportion of G1 in NP69 - LMP1^△232-351 were more than in NP69 - LMP1^WT ( p 〈 0.01 ). Conclusion Carboxy terminal activating region - 3 of LMPI probably mediated to regulate cell cycle and apoptosis to affect biological character in nasopharyngeal epithelial cell.